Structure of Mitochondrial Channel VDAC: Insights from Site-Directed Mutagenesis

1. The figures in this presentation were taken from: http://www.wadsworth.org/ Blachly-Dyson, E., Peng, S. Z., Colombini, M. and Forte, M. 1989. Probing the structure of the mitochondrial channel, VDAC, by site-directed mutagenesis: a progress report. Journal of Bioenergetics and Biomembranes, 21:471-483. Blachly-Dyson, E., Peng, S. Z., Colombini, M. and Forte, M. 1990. Alteration of the selectivity of the VDAC ion channel by site-directed mutagenesis: implications for the structure of a membrane ion channel. Science, 247: 1233-1236. Thomas, L., Kocsis, E., Colombini, M., Erbe, E., Trus, B.L. and Steven, A.C. 1991. Surface topography and molecular stoichiometry of the mitochondrial channel, VDAC, in crystalline arrays. Journal of Structural Biology, 106: 161-171. Peng, S., Blachly-Dyson, E., Colombini, M. and Forte, M. 1992. Large scale rearrangement of protein domains is associated with voltage gating of the VDAC channel. Biophysical Journal, 62: 123-153. Thomas, L., Blachly-Dyson, E., Colombini, M., and Forte, M. 1993. Mapping of residues forming the voltage sensor of the VDAC ion channel. Proceedings of the National Academy of Sciences U.S.A., 90: 5446-5449. Song, J., Midson, C., Blachly-Dyson, E., Forte, M., and Colombini, M. 1998. The sensor regions of VDAC are translocated from within the membrane to the surface during the gating processes. Biophysical Journal, 74: 2926-2944. Song, J., Midson, C., Blachly-Dyson, E., Forte, M., and Colombini, M. 1998. The topology of VDAC as probed by biotin modification. Journal of Biological Chemistry, 273:24406‑24413. Vander Heiden, M.G., Chandel, N.S., Li, X.X., Schumacker, P.T., Colombini, M., and Thompson, C.B. 2000. Outer mitochondrial membrane permeability can regulate coupled respiration and cell survival. Proceedings of the National Academy of Sciences U.S.A., 97: 4666‑4671.

2. a mitochondrion... work of Carmen Mannella from: http://www.wadsworth.org/BMS/SCBlinks/channels2.htm http://www.wadsworth.org/rvbc/tomography.html

3. Images of VDAC Biophysics of Channels Associated with Apoptosis reconstruction of negatively-stained image by Carmen Mannella from: http://www.wadsworth.org /BMS/SCBlinks/vdac.htm 2D-crystals in mitochon-drial outer membrane averaged surface view

5. A N K I Y Q K V N K K L E T A V N L S K I F T A T E K K G W L K G T G K A A I G F R T N S N G A T G W T G F Y Y N N Y E Q E T P K T I A V P P G T D S R G D K T N L W N F E Y E D P L P A Y A E E R T S H G G N D T G Q D L S F Y L S I L W V A Q I F G K K T K G D N V L N C T K E S A T S T T F L L A T F Y L L R D K S E E T G A G H S G T G V F L G L I L C G Y L A L L L T K D S S T K D R Q Q K G S G G Y L A G V L M I M F V I A L G F K N T E G D S N E N L L K G I T V D R F P F D N S L H L E K Q A D G E T S D G L I A T T T K G G A Y K A V R S K G N N T V Q S N F A V K111 A F G N S Y Y H K V N S Q V E A G S K A A R L T A H S189 W N L T68 N F F K T A V E L G V T N G T K S G V T K V Y N T A Q S R P Q P S L I E252 A V P N37 K T L N N D R G A N V D W F F H E144 D P L V S282 F S P A T N S H G G T V L T E D I T F F T F G V F A S V L F A K N K K A I R I L T F N G T S A S V G N G A A A I V Y V F N D K T E A E F D G A K A G S L L V G L L A F I A A G I A T N K E K A E K D N S S K A S G D F I S G T L L A A Y I A F V Y H T T S K G L T G T N V D K L G H T V K K P Y P D G T H A G A D V E A D A R Q T K M L V H K A A N I A A K Q Y L Q D D T G G Y S A A V human VDAC1 N. crassa VDAC

7. VDAC Selectivity Change Upon Closure 2-

8. applied voltage current recording

21. (25 mM)

26. Biotin modification + Avidin binding biotin